CN101700368A - Method for detecting components of pharmaceutical composition for treating diseases of urinary system - Google Patents

Method for detecting components of pharmaceutical composition for treating diseases of urinary system Download PDF

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CN101700368A
CN101700368A CN200910224024A CN200910224024A CN101700368A CN 101700368 A CN101700368 A CN 101700368A CN 200910224024 A CN200910224024 A CN 200910224024A CN 200910224024 A CN200910224024 A CN 200910224024A CN 101700368 A CN101700368 A CN 101700368A
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herba
poria
water
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acid
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CN101700368B (en
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何彦杰
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Jiangxi Hongxing Pharmaceutical Co Ltd
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Jiangxi Hongxing Pharmaceutical Co Ltd
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Abstract

The invention relates to a method for detecting components of a pharmaceutical composition for treating diseases of the urinary system, which comprises the following steps: taking the pharmaceutical composition as the sample solution; making a herba plantaginis medicine into a control medicine solution; and absorbing the two solutions, respectively dropping on the same silica gel G thin-layer plate using sodium carboxymethyl cellulose as the adhesive, developing by using ethyl acetate-methanoic acid-water as the developing agent, taking out, drying in the air, spraying a vanillina-sulphuric acid solution, heating until the spots become clear, and putting in a sample chromatogram, wherein in the positions corresponding to the control medicine chromatogram, the spots with the same colors are displayed. The invention has the advantages of low cost of preparation technique and detecting technique, and easy processing, reduces the heating time of the medicine and is suitable for wide application in clinical treatment.

Description

The detection method of the components of pharmaceutical composition of treatment diseases of urinary system
Technical field
The present invention relates to a kind of detection method for the treatment of the components of pharmaceutical composition of diseases of urinary system.
Background technology
Diseases of urinary system is human common disease, its cardinal symptom have frequent micturition short red, urinate puckery causalgia, serious occurred heating, thirsty, abdominal pain, waist is ached, hematuria, suppurate, constipation with dry stool or completely mashed.Urinary system calculus belongs to the stranguria caused by urinary stone in the Chinese medicine stranguria.Chinese medicine is thought by damp invasion of lower energizer, and turning into fire and then burning YIN decocts urine, becomes sandstone, and the alluvial water channel forms.Dialectical excess syndrome based on part of the body cavity below the umbilicus, housing the bladder, kidneys and bowels dampness and heat stasis.Control suitable clearing away heat-damp and promoting diuresis, Tonglin Paishi.So get rid of the key that the water channel sandstone will become the treatment urinary system calculus.At present the western medicine therapy to kidney, urinary tract, wing skin calculus is surgical calculus removing and external instrument concussion rubble calculus method, operates on modus operandi expense height, patient's misery and postoperative recurrence rate height; Though it is fast that calculus method speed is impacted in instrument concussion, short treating period, the equipment manufacturing cost height is not suitable for the extensive patients in samll cities and towns or rural area.The traditional Chinese medical science then is to adopt various calculus prescriptions, and its cost is lower, but most prescription among the people does not pass through strict pharmacodynamic experiment and toxicity test, and also each is variant for therapeutic effect; To acute pyelonephritis, wing skin inflammation, the Therapeutic Method of the present doctor trained in Western medicine of disease that urethritis and sexually transmitted disease (STD) cause adopts antibiotic therapy mostly, the therapeutic effect of antibiotics is obvious, speed is fast, but it is not remarkable to the chronic disease therapeutic effect, the medication that has also can produce the drug-resistant effect to antibacterial too for a long time, so the traditional Chinese medical science has been studied some preparations at present, in order to treatment urinary system calculus and urinary tract infection, as SANJIN PIAN (capsule), the treating stranguria oral medicine of money, BAZHENG HEJI, JISHENG SHENQI WAN, composite shi-wei tablet etc., these Chinese medicine preparation have certain therapeutical effect to diseases such as treatment urinary system calculus and urinary tract infection, but do not obtain satisfactory effect yet.
Summary of the invention
The present invention provides a kind of detection method for the treatment of the components of pharmaceutical composition of diseases of urinary system just from prior art, thereby its cost is reduced, and handling ease is suitable in the extensive use clinical treatment.
The detection method of the components of pharmaceutical composition of treatment diseases of urinary system of the present invention, the composition of the pharmaceutical composition of wherein said treatment diseases of urinary system is Herba Desmodii Styracifolii, Stigma Maydis, Folium Pyrrosiae, Herba Abri, Poria, Herba Plantaginis, Herba lygodii, Rhizoma Imperatae, and each composition percentage by weight is as follows:
Herba Desmodii Styracifolii 21～24%
Stigma Maydis 12～18%
Folium Pyrrosiae 9～13%
Herba Abri 8～13%
Poria 8～12%;
Herba Plantaginis 7～10%
Herba lygodii 7～10%
Rhizoma Imperatae 8～13%
And chlorogenic acid (C in the described Folium Pyrrosiae 16H 18O 9) content, must not be less than 0.3% of gross weight, above-mentioned medical material is obtained described pharmaceutical composition by following method for making:
Step 1: get described Poria and be divided into two parts, it is 80～90% that a copy of it accounts for the Poria total weight percent, and it is 10～20% that another part accounts for the Poria total weight percent;
Step 2: will account for the Poria total weight percent and be 80～90% portion and be ground into fine powder;
Step 3: will remain Poria and all the other seven flavor medicine materials decoct with water 1～3 time, each 2～4 hours, collecting decoction filtered, and filtrate is condensed into the clear paste that relative density is 1.19～1.20 (90 ℃);
Step 4: add the Poria fine powder, mixing, drying is ground into fine powder, sieves, and adds appropriate amount of auxiliary materials, and mixing is granulated drying, granulate;
Described detection method comprises:
Compositions 5～15g gets it filled, add water 30～70ml and make dissolving, in 60～90 ℃ of extractions 1～3 time, each 30～50ml discards petroleum ether liquid with petroleum ether, ethyl acetate extraction 1～3 time of water liquid, each 20～40ml merges ethyl acetate liquid, evaporate to dryness, residue adds ethanol 1～3ml makes dissolving, as need testing solution;
Other gets Herba Plantaginis control medicinal material 1～3g, adds 70% ethanol, 100～300ml, refluxes 2～3 hours, puts coldly, filters, and filtrate evaporate to dryness, residue add water 30～50ml makes dissolving, extracts 2～4 times for 60～90 ℃ with petroleum ether, gets control medicinal material solution with legal system;
Draw above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, with ethyl acetate-formic acid-water is developing solvent, launches, and takes out, dry, spray is heated to clear spot with the vanillin sulfuric acid solution, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
Preferably, the compositions 10g that gets it filled adds water 50ml and makes dissolving, extracts 2 times for 60～90 ℃ with petroleum ether, each 40ml discards petroleum ether liquid, ethyl acetate extraction 2 times of water liquid, each 30ml, merge ethyl acetate liquid, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution;
Other gets Herba Plantaginis control medicinal material 1g, adds 70% ethanol 100ml, refluxes 2 hours, puts coldly, filters, and filtrate evaporate to dryness, residue add water 30ml makes dissolving, extracts 2 times with 60～90 ℃ of petroleum ether and rises, and gets control medicinal material solution with legal system;
Draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, with ethyl acetate-formic acid of 6: 1: 1-water is developing solvent, launches, and takes out, dry, spray is heated to clear spot at 105 ℃, in the test sample chromatograph with 5% vanillin sulfuric acid solution, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
On the other hand, detection method also comprises chlorogenic acid (C 16H 18O 9) the step of assay, described assay adopts high effective liquid chromatography for measuring, condition determination is as follows:
Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Acetonitrile-0.5% phosphoric acid solution (11: 89) is a mobile phase; The detection wavelength is 326nm, and number of theoretical plate calculates by the chlorogenic acid peak should be not less than 3000;
It is an amount of that the preparation precision of reference substance solution takes by weighing the chlorogenic acid reference substance, adds methanol and make the solution that every 1ml contains 60 μ g, promptly;
The about 1.0g of this product content under the content uniformity item is got in the preparation of need testing solution, the accurate title, decide, and puts in the 100ml round-bottomed flask, added the 50ml methanol eddy 2 hours, filter, with about 20ml gradation washing medicinal residues of methanol and filter, merging filtrate, evaporate to dryness, residue add methanol make in right amount the dissolving and be transferred in the 10ml measuring bottle, add methanol to scale, shake up, promptly;
Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly.
Preferably, wherein said Herba Desmodii Styracifolii main component is soybean saponin I, puriri glycosides-1, puriri glycosides-3, Xia Futa basket glycosides, described Stigma Maydis main component is a fatty oil, volatile oil, alkaloid, ascorbic acid, sitosterol, described Folium Pyrrosiae main component is the bracken triterpene, cupreol, kaempferol, Quercetin, isoquercitrin, Trifolin, described Herba Abri main component is a triterpenes: sophoradiol, abrisapogenol A-G. glycyrrhizin, described Poria main component is β-pachyman, the acetyl pachymic acid, pachymic acid, chitin, protein, choline, described Herba Plantaginis main component is an aucubin, Homoplantaginin, ursolic acid, cupreol, described Herba lygodii main component is an aminoacid, saccharide, flavonoid glycoside and phenols, described Rhizoma Imperatae main component is an arundoin, the Yin Baimao element, Isoarborinol alcohol.
Preferably, amount of water is 10～12 times of amounts of medical material weight in the step 3.
Preferred, amount of water is 10 times of amounts of medical material weight in the step 3.
Preferred, decoct with water secondary in the step 3, each 3 hours, collecting decoction filtered, and filtrate is concentrated into the clear paste that relative density is 1.20 (90 ℃).
Preferred, wherein the optimum weight percentage ratio of each composition is:
Herba Desmodii Styracifolii 22.8%
Stigma Maydis 15.1%
Folium Pyrrosiae 11.4%
Herba Abri 11.4%
Poria 11.4%
Herba Plantaginis 8.25%
Herba lygodii 8.25%
Rhizoma Imperatae 11.4%
Preparation technology of the present invention and characterization processes cost are low, and handling ease reduces the medicine heated time, is suitable in the extensive use clinical treatment.Adopt the pharmaceutical composition of the inventive method preparation, have broad-spectrum antiseptic, bacteriostasis, significant diuresis is arranged and regulate the effect of ureter smooth muscle.And have the effect of clearing heat and expelling damp, anti-inflammatory analgetic, diuresis and expelling stone, and can be used for treating diseases such as urinary system calculus and urinary tract infection, have diuresis simultaneously, make the urine dilution, can reduce crystal formation., urinary tract infection damp and hot for the wing skin, lithangiuria, renal colic etc. have good auxiliary treatment effect.
Description of drawings
The canonical plotting that Fig. 1 chlorogenic acid is measured
The specific embodiment
Below, provide some specific embodiments that the present invention treats the preparation of pharmaceutical compositions of diseases of urinary system.
Embodiment 1
Herba Desmodii Styracifolii 285g Stigma Maydis 188.8g Folium Pyrrosiae 142.5g
Herba Abri 142.5g Poria 142.5g Herba Plantaginis 103.1g
Herba lygodii 103.1g Rhizoma Imperatae 142.5g
Folium Pyrrosiae chlorogenic acid (C 16H 18O 9) content 15g
[method for making] above eight flavor medicines are got Poria 120g and are ground into fine powder, and are standby; Residue Poria and Herba Desmodii Styracifolii, Stigma Maydis, Folium Pyrrosiae, Herba Abri, Herba Plantaginis, Herba lygodii and the merging of Rhizoma Imperatae seven flavor medicine decoct with water twice, and each 3 hours, merging filtrate, filter, filtrate is concentrated into the extractum of 1.19-1.20 (90 ℃), adds the Poria fine powder, mix homogeneously is dried to dried cream, is ground into fine powder, sieve, add appropriate amount of auxiliary materials, mixing, granulate drying, granulate, incapsulate, make 1000, promptly.
Embodiment 2
Herba Desmodii Styracifolii 285g Stigma Maydis 188.8g Folium Pyrrosiae 187.5g
Herba Abri 97.5g Poria 142.5g Herba Plantaginis 103.1g
Herba lygodii 103.1g Rhizoma Imperatae 142.5g
Folium Pyrrosiae chlorogenic acid (C 16H 18O 9) content 20g
[method for making] above eight flavor medicines are got Poria 120g and are ground into fine powder, and are standby; Residue Poria and Herba Desmodii Styracifolii, Stigma Maydis, Folium Pyrrosiae, Herba Abri, Herba Plantaginis, Herba lygodii and the merging of Rhizoma Imperatae seven flavor medicine decoct with water twice, and each 3 hours, merging filtrate, filter, filtrate is concentrated into the extractum of 1.19-1.20 (90 ℃), adds the Poria fine powder, mix homogeneously is dried to dried cream, is ground into fine powder, sieve, add appropriate amount of auxiliary materials, mixing, granulate drying, granulate, incapsulate, make 1000, promptly.
Embodiment 3
Herba Desmodii Styracifolii 285g Stigma Maydis 188.8g Folium Pyrrosiae 100g
Herba Abri 260g Poria 142.5g Herba Plantaginis 103.1g
Herba lygodii 103.1g Rhizoma Imperatae 142.5g
Folium Pyrrosiae chlorogenic acid (C 16H 18O 9) content 10g
[method for making] above eight flavor medicines are got Poria 120g and are ground into fine powder, and are standby; Residue Poria and Herba Desmodii Styracifolii, Stigma Maydis, Folium Pyrrosiae, Herba Abri, Herba Plantaginis, Herba lygodii and the merging of Rhizoma Imperatae seven flavor medicine decoct with water twice, and each 3 hours, merging filtrate, filter, filtrate is concentrated into the extractum of 1.19-1.20 (90 ℃), adds the Poria fine powder, mix homogeneously is dried to dried cream, is ground into fine powder, sieve, add appropriate amount of auxiliary materials, mixing, granulate drying, granulate, incapsulate, make 1000, promptly.
Embodiment 4
Herba Desmodii Styracifolii 285g Stigma Maydis 188.8g Folium Pyrrosiae 222.5g
Herba Abri 62.5g Poria 142.5g Herba Plantaginis 103.1g
Herba lygodii 103.1g Rhizoma Imperatae 142.5g
Folium Pyrrosiae chlorogenic acid (C 16H 18O 9) content 23g
[method for making] above eight flavor medicines are got Poria 120g and are ground into fine powder, and are standby; Residue Poria and Herba Desmodii Styracifolii, Stigma Maydis, Folium Pyrrosiae, Herba Abri, Herba Plantaginis, Herba lygodii and the merging of Rhizoma Imperatae seven flavor medicine decoct with water twice, and each 3 hours, merging filtrate, filter, filtrate is concentrated into the extractum of 1.19-1.20 (90 ℃), adds the Poria fine powder, mix homogeneously is dried to dried cream, is ground into fine powder, sieve, add appropriate amount of auxiliary materials, mixing, granulate drying, granulate, incapsulate, make 1000, promptly.
Embodiment 5
Herba Desmodii Styracifolii 285g Stigma Maydis 188.8g Folium Pyrrosiae 135g
Herba Abri 142.5g Poria 150g Herba Plantaginis 103.1g
Herba lygodii 103.1g Rhizoma Imperatae 142.5g
Folium Pyrrosiae chlorogenic acid (C 16H 18O 9) content 14g
[method for making] above eight flavor medicines are got Poria 130g and are ground into fine powder, and are standby; Residue Poria and Herba Desmodii Styracifolii, Stigma Maydis, Folium Pyrrosiae, Herba Abri, Herba Plantaginis, Herba lygodii and the merging of Rhizoma Imperatae seven flavor medicine decoct with water twice, and each 3 hours, merging filtrate, filter, filtrate is concentrated into the extractum of 1.19-1.20 (90 ℃), adds the Poria fine powder, mix homogeneously is dried to dried cream, is ground into fine powder, sieve, add appropriate amount of auxiliary materials, mixing, granulate drying, granulate, incapsulate, make 1000, promptly.
Embodiment 6
Herba Desmodii Styracifolii 312.5g Stigma Maydis 188.8g Folium Pyrrosiae 115g
Herba Abri 142.5g Poria 142.5g Herba Plantaginis 103.1g
Herba lygodii 103.1g Rhizoma Imperatae 142.5g
Folium Pyrrosiae chlorogenic acid (C 16H 18O 9) content 12g
[method for making] above eight flavor medicines are got Poria 120g and are ground into fine powder, and are standby; Residue Poria and Herba Desmodii Styracifolii, Stigma Maydis, Folium Pyrrosiae, Herba Abri, Herba Plantaginis, Herba lygodii and the merging of Rhizoma Imperatae seven flavor medicine decoct with water twice, and each 3 hours, merging filtrate, filter, filtrate is concentrated into the extractum of 1.19-1.20 (90 ℃), adds the Poria fine powder, mix homogeneously is dried to dried cream, is ground into fine powder, sieve, add appropriate amount of auxiliary materials, mixing, granulate drying, granulate, incapsulate, make 1000, promptly.
Embodiment 7
Herba Desmodii Styracifolii 312.5g Stigma Maydis 62.5g Folium Pyrrosiae 268.8g
Herba Abri 142.5g Poria 142.5g Herba Plantaginis 103.1g
Herba lygodii 103.1g Rhizoma Imperatae 142.5g
Folium Pyrrosiae chlorogenic acid (C 16H 18O 9) content 28g
[method for making] above eight flavor medicines are got Poria 120g and are ground into fine powder, and are standby; Residue Poria and Herba Desmodii Styracifolii, Stigma Maydis, Folium Pyrrosiae, Herba Abri, Herba Plantaginis, Herba lygodii and the merging of Rhizoma Imperatae seven flavor medicine decoct with water twice, and each 3 hours, merging filtrate, filter, filtrate is concentrated into the extractum of 1.19-1.20 (90 ℃), adds the Poria fine powder, mix homogeneously is dried to dried cream, is ground into fine powder, sieve, add appropriate amount of auxiliary materials, mixing, granulate drying, granulate, incapsulate, make 1000, promptly.
Embodiment 8
Herba Desmodii Styracifolii 162.5g Stigma Maydis 188.8g Folium Pyrrosiae 158.8g
Herba Abri 142.5g Poria 142.5g Herba Plantaginis 125g
Herba lygodii 87.5g Rhizoma Imperatae 142.5g
Folium Pyrrosiae chlorogenic acid (C 16H 18O 9) content 16g
[method for making] above eight flavor medicines are got Poria 133g and are ground into fine powder, and are standby; Residue Poria and Herba Desmodii Styracifolii, Stigma Maydis, Folium Pyrrosiae, Herba Abri, Herba Plantaginis, Herba lygodii and the merging of Rhizoma Imperatae seven flavor medicine decoct with water twice, and each 3 hours, merging filtrate, filter, filtrate is concentrated into the extractum of 1.19-1.20 (90 ℃), adds the Poria fine powder, mix homogeneously is dried to dried cream, is ground into fine powder, sieve, add appropriate amount of auxiliary materials, mixing, granulate drying, granulate, incapsulate, make 1000, promptly.
Embodiment 9
Herba Desmodii Styracifolii 300g Stigma Maydis 150g Folium Pyrrosiae 142.5g
Herba Abri 142.5g Poria 142.5g Herba Plantaginis 105g
Herba lygodii 125g Rhizoma Imperatae 142.5g
Folium Pyrrosiae chlorogenic acid (C 16H 18O 9) content 15g
[method for making] above eight flavor medicines are got Poria 120g and are ground into fine powder, and are standby; Residue Poria and Herba Desmodii Styracifolii, Stigma Maydis, Folium Pyrrosiae, Herba Abri, Herba Plantaginis, Herba lygodii and the merging of Rhizoma Imperatae seven flavor medicine decoct with water twice, and each 3 hours, merging filtrate, filter, filtrate is concentrated into the extractum of 1.19-1.20 (90 ℃), adds the Poria fine powder, mix homogeneously is dried to dried cream, is ground into fine powder, sieve, add appropriate amount of auxiliary materials, mixing, granulate drying, granulate, incapsulate, make 1000, promptly.
Embodiment 10
Herba Desmodii Styracifolii 291.3g Stigma Maydis 225g Folium Pyrrosiae 142.5g
Herba Abri 100g Poria 142.5g Herba Plantaginis 103.1g
Herba lygodii 103.1g Rhizoma Imperatae 142.5g
Green former (the C of Folium Pyrrosiae 16H 18O 9) content 15g
[method for making] above eight flavor medicines are got Poria 120g and are ground into fine powder, and are standby; Residue Poria and Herba Desmodii Styracifolii, Stigma Maydis, Folium Pyrrosiae, Herba Abri, Herba Plantaginis, Herba lygodii and the merging of Rhizoma Imperatae seven flavor medicine decoct with water twice, and each 3 hours, merging filtrate, filter, filtrate is concentrated into the extractum of 1.19-1.20 (90 ℃), adds the Poria fine powder, mix homogeneously is dried to dried cream, is ground into fine powder, sieve, add appropriate amount of auxiliary materials, mixing, granulate drying, granulate, incapsulate, make 1000, promptly.
Embodiment 11
Herba Desmodii Styracifolii 300g Stigma Maydis 218.8g Folium Pyrrosiae 162.5g
Herba Abri 162.5g Poria 100g Herba Plantaginis 103.1g
Herba lygodii 103.1g Rhizoma Imperatae 100g
Folium Pyrrosiae chlorogenic acid (C 16H 18O 9) content 17g
[method for making] above eight flavor medicines are got Poria 86g and are ground into fine powder, and are standby; Residue Poria and Herba Desmodii Styracifolii, Stigma Maydis, Folium Pyrrosiae, Herba Abri, Herba Plantaginis, Herba lygodii and the merging of Rhizoma Imperatae seven flavor medicine decoct with water twice, and each 3 hours, merging filtrate, filter, filtrate is concentrated into the extractum of 1.19-1.20 (90 ℃), adds the Poria fine powder, mix homogeneously is dried to dried cream, is ground into fine powder, sieve, add appropriate amount of auxiliary materials, mixing, granulate drying, granulate, incapsulate, make 1000, promptly.
Embodiment 12
Herba Desmodii Styracifolii 285g Stigma Maydis 223.8g Folium Pyrrosiae 142.5g
Herba Abri 142.5g Poria 150g Herba Plantaginis 103.1g
Herba lygodii 103.1g Rhizoma Imperatae 100g
Folium Pyrrosiae chlorogenic acid (C 16H 18O 9) content 15g
[method for making] above eight flavor medicines are got Poria 120g and are ground into fine powder, and are standby; Residue Poria and Herba Desmodii Styracifolii, Stigma Maydis, Folium Pyrrosiae, Herba Abri, Herba Plantaginis, Herba lygodii and the merging of Rhizoma Imperatae seven flavor medicine decoct with water twice, and each 3 hours, merging filtrate, filter, filtrate is concentrated into the extractum of 1.19-1.20 (90 ℃), adds the Poria fine powder, mix homogeneously is dried to dried cream, is ground into fine powder, sieve, add appropriate amount of auxiliary materials, mixing, granulate drying, granulate, incapsulate, make 1000, promptly.
Embodiment 13
Herba Desmodii Styracifolii 285g Stigma Maydis 188.8g Folium Pyrrosiae 112.5g
Herba Abri 142.5g Poria 142.5g Herba Plantaginis 103.1g
Herba lygodii 113.1g Rhizoma Imperatae 162.5g
Folium Pyrrosiae chlorogenic acid (C 16H 18O 9) content 11g
[method for making] above eight flavor medicines are got Poria 95g and are ground into fine powder, and are standby; Residue Poria and Herba Desmodii Styracifolii, Stigma Maydis, Folium Pyrrosiae, Herba Abri, Herba Plantaginis, Herba lygodii and the merging of Rhizoma Imperatae seven flavor medicine decoct with water twice, and each 3 hours, merging filtrate, filter, filtrate is concentrated into the extractum of 1.19-1.20 (90 ℃), adds the Poria fine powder, mix homogeneously is dried to dried cream, is ground into fine powder, sieve, add appropriate amount of auxiliary materials, mixing, granulate drying, granulate, incapsulate, make 1000, promptly.
The present invention treat the percentage by weight of each composition of pharmaceutical composition preferred implementation of diseases of urinary system can following subordinate list 1 shown in:
Subordinate list 1:
Herba Desmodii Styracifolii ??22.80％ ??21.00％ ??24.00％ ??23.30％ ??24.00％ ??22.80％ ??22.80％
Stigma Maydis ??15.10％ ??15.10％ ??12.00％ ??18.00％ ??17.50％ ??17.90％ ??15.10％
Folium Pyrrosiae ??11.40％ ??12.70％ ??11.40％ ??11.40％ ??13.00％ ??11.40％ ??11.40％
Herba Abri ??11.40％ ??11.40％ ??11.40％ ??8.00％ ??13.00％ ??11.40％ ??11.40％
Poria ??11.40％ ??11.40％ ??11.40％ ??11.40％ ??8.00％ ??12.00％ ??11.40％
Herba Desmodii Styracifolii ??22.80％ ??21.00％ ??24.00％ ??23.30％ ??24.00％ ??22.80％ ??22.80％
Herba Plantaginis ??8.25％ ??10.00％ ??8.40％ ??8.25％ ??8.25％ ??8.25％ ??7.00％
Herba lygodii ??8.25％ ??7.00％ ??10.00％ ??8.25％ ??8.25％ ??8.25％ ??7.90％
Rhizoma Imperatae ??11.40％ ??11.40％ ??11.40％ ??11.40％ ??8.00％ ??8.00％ ??13.00％
In addition,, further provide the detection method of its component, provide some specific embodiments below in conjunction with the pharmaceutical composition of treatment diseases of urinary system of the present invention, as follows:
Embodiment 14
Get this product 5g, add water 30ml and make dissolving, extract 1 time, each 30ml with petroleum ether (60～90 ℃), discard petroleum ether liquid, ethyl acetate extraction 1 time of water liquid, each 30ml merges ethyl acetate liquid, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution.Other gets Herba Plantaginis control medicinal material 1g, adds 70% ethanol 100ml, refluxes 2 hours, puts coldly, filters, and filtrate evaporate to dryness, residue add water 30ml makes dissolving, and personal petroleum ether (60～90 ℃) extracts 2 times and rises, and gets control medicinal material solution with legal system.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, be developing solvent with ethyl acetate-formic acid-water (6: 1: 1), launch, take out, dry, spray is with 5% vanillin sulfuric acid solution, be heated to clear spot at 105 ℃, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
Embodiment 15
Get this product 10g, add water 50ml and make dissolving, extract 2 times, each 40ml with petroleum ether (60～90 ℃), discard petroleum ether liquid, ethyl acetate extraction 2 times of water liquid, each 30ml merges ethyl acetate liquid, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution.Other gets Herba Plantaginis control medicinal material 1g, adds 70% ethanol 100ml, refluxes 2 hours, puts coldly, filters, and filtrate evaporate to dryness, residue add water 30ml makes dissolving, and personal petroleum ether (60～90 ℃) extracts 2 times and rises, and gets control medicinal material solution with legal system.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, be developing solvent with ethyl acetate-formic acid-water (6: 1: 1), launch, take out, dry, spray is with 5% vanillin sulfuric acid solution, be heated to clear spot at 105 ℃, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
Embodiment 16
Get this product 15g, add water 70ml and make dissolving, extract 3 times, each 50ml with petroleum ether (60～90 ℃), discard petroleum ether liquid, ethyl acetate extraction 3 times of water liquid, each 40ml merges ethyl acetate liquid, evaporate to dryness, residue add ethanol 3ml makes dissolving, as need testing solution.Other gets Herba Plantaginis control medicinal material 3g, adds 70% ethanol 300ml, refluxes 3 hours, puts coldly, filters, and filtrate evaporate to dryness, residue add water 30ml makes dissolving, and personal petroleum ether (60～90 ℃) extracts 4 times, gets control medicinal material solution with legal system.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, be developing solvent with ethyl acetate-formic acid-water (6: 1: 1), launch, take out, dry, spray is with 5% vanillin sulfuric acid solution, be heated to clear spot at 105 ℃, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
Simultaneously, the present invention also provides a kind of method of Folium Pyrrosiae medical material determination of chlorogenic acid and related assays result to show that embodiment is as follows:
1, instrument and reagent
American SS I PC2000 type high performance liquid chromatograph, 500 type UV, visible light detectors; The ShimpackC18 post (6.0 * 150mm), the Anastar chromatographic work station;
Germany's BP211D Sartorius electronic balance (100,000/);
The clear device of JL-120 type ultrasound wave;
Chlorogenic acid reference substance: provide lot number by Nat'l Pharmaceutical ﹠ Biological Products Control Institute: 110753-200212
Agents useful for same is analytical pure or chromatographically pure.
2, the selection of extraction separation method
Physicochemical property according to chlorogenic acid, adopt methanol ultrasonic method and methanol eddy legal system to be equipped with need testing solution respectively, through injecting hplc determination, methanol content that ultrasonic method is surveyed is 0.148% as a result, circumfluence method is 0.265%, so select the methanol eddy legal system to be equipped with test liquid, carried out the tracking of return time simultaneously, the results are shown in following table.
The tracking of table 1 reflux extracting time
Figure G2009102240244D0000121
By above-mentioned experimental result as can be known, reflux, extract, can be extracted chlorogenic acid fully in 4 hours substantially.
3, the preparation of need testing solution
Get the about 0.5g of this product coarse powder, accurate claim surely, put in the 100ml round-bottomed flask, added the 50ml methanol eddy 4 hours, filter, the methanol solution water-bath volatilizes, and residue adds dissolve with methanol and standardize solution in the 25ml measuring bottle, shakes up, promptly.
4, the preparation precision of reference substance solution takes by weighing chlorogenic acid reference substance 2.95mg, puts in the 50ml measuring bottle, adds methanol ultrasonic dissolution and standardize solution in scale, shakes up, promptly.
5, the selection of chromatographic condition
Adopting acetonitrile-0.5% phosphoric acid solution (11: 89) is mobile phase; The detection wavelength is 326nm, and flow velocity is 1.0ml/min, and column temperature is 30 ℃, under this chromatographic condition, chlorogenic acid peak energy and other peaks reach baseline separation in the test sample chromatograph, and peak shape is better, consistent with chlorogenic acid reference substance peak retention time, relevant HPLC chromatogram is seen Fig. 5.
6, chromatographic condition and system suitability test
The chromatographic condition octadecylsilane chemically bonded silica is a filler, and acetonitrile-0.5% phosphoric acid solution (11: 89) is a mobile phase; The detection wavelength is 326nm, flow velocity: 1.0ml/min, column temperature: 30 ℃.
The accurate test liquid 10 μ l that draw of system suitability test inject chromatograph of liquid, measure, and press the chlorogenic acid peak and calculate number of theoretical plate n=3014, tailing factor T=1.037, separating degree R=5.329.
The accurate chlorogenic acid reference substance solution 10 μ l that draw of replica test inject chromatograph of liquid, and continuous sample introduction 5 times calculates RSD, the results are shown in following table.
Table 2 replica test
7, the investigation of the preparation of standard curve and linear relationship
Accurate reference substance solution (59 μ g/ml) 2.5,5.0,7.5,10.0, the 12.5 and 15 μ l that draw inject chromatograph of liquid, measure, with the chlorogenic acid amount is abscissa, the chlorogenic acid peak area is a vertical coordinate, drawing standard curve (see figure 1), experimental data returns through linearity, the standard curve equation is: Y=2048948.184X-28108.333r=0.99997 (n=6), chlorogenic acid reference substance amount is good linear relationship with its peak area in 0.1475 μ g～0.8850 μ g scope, the results are shown in following table.
The preparation of table 3 standard curve
Figure G2009102240244D0000132
8, precision test
The accurate need testing solution 10 μ l that draw inject chromatograph of liquid, repeat sample introduction 5 times, the results are shown in following table.
The test of table 4 precision
Figure G2009102240244D0000133
9, stability test
Get need testing solution, the 1 hour sample introduction at room temperature every interval is once measured peak area, calculates RSD, and experiment shows that chlorogenic acid is basicly stable in 4 hours of being surveyed, and the results are shown in following table.
Table 5 stability test
10, repeatability test
Get with 5 parts of batch medical materials, measure, the results are shown in following table by the method under the text assay item.
The test of table 6 repeatability
Figure G2009102240244D0000142
11, average recovery test
Get 5 parts of this product coarse powder (content is 0.274%) of known content, accurately claim surely, the accurate respectively chlorogenic acid reference substance that adds is an amount of, presses the method mensuration text assay item under, the results are shown in following table.
Table 7 average recovery result of the test
Figure G2009102240244D0000143
The clinical observation curative effect:
The calculus open capsule is a kind of Chinese patent medicine that Herba Desmodii Styracifolii, Folium Pyrrosiae, Poria, Herba Plantaginis, Spora Lygodii etc. are made, the clinical observation curative effect of its treatment urinary system calculus;
Therapeutic scheme: 180 examples are divided into treatment at random and organize 130 examples, oral calculus open capsule (producing) by Jiangxi Red Star pharmaceutcal corporation, Ltd, and lot number: (20060701), each 5, every day 3 times, oral.Matched group 50 examples, each 5 of oral SHILINTONG PIAN (manufacturing the production of medicine company limited in Zhanjiang), every day 3 times, oral.
The course of treatment and other.20 is 1 course of treatment, and state of an illness no change continues to observe second course of treatment.Advise the patient to drink water more than 1500ml every day during the treatment, do jumping exercise more.
Observational technique: all objects of observation are all at treatment before and after look routine blood test, and routine urinalysis, liver, renal function, B ultrasonic, plain abdominal radiograph are observed drug side effect such as indigestion and loss of appetite, abdominal distention and pain, nausea and vomiting, dizzy, headache etc., and liver, renal function injury.
Criterion of therapeutical effect, cure: calculus is discharged, transference cure.B ultrasonic or KUB check calculus disappear; Take a turn for the better: doing well,improving, B ultrasonic or KUB check calculus is dwindled or the position moves down; Invalid: symptom and check B ultrasonic or KUB calculus do not have change.
The result:
12 groups of curative effects relatively
Group ?n Cure Invalid Total effective rate %
The treatment group ?130 ??81 ??35 ??89.2
Matched group ?50 ??26 ??12 ??76.0
Group is imitated relatively, P＜0.05
22 groups of symptoms relatively
Figure G2009102240244D0000151
Figure G2009102240244D0000161
Compare P＜0.05 with matched group
2 groups of each position calculus curative effects relatively
Figure G2009102240244D0000162
Compare P＜0.05 with matched group
By 2 groups of clinical test results relatively, calculus open capsule treatment group cure rate 62.3%, total effective rate 89.2%, matched group is respectively 52.0%, 76.0%, 2 group more all significant difference (P＜0.05), calculus open capsule, removing damp-heat is arranged, Tonglin Paishi, the significant curative effect of pain easing and hemostasis is simultaneously at infection with improve and significant curative effect is also arranged aspect the renal function.
Calculus open capsule toxicological experiment is as follows:
Sample is capsule from Jiangxi Red Star pharmaceutcal corporation, Ltd, and content is tan powder, every 0.35g, and the oral recommended amounts of human body is 12 of every days, becomes body weight for humans to press 60kg and calculates, and amounts to dosage 0.07g/kg.Acute toxicity test in mice adopts mtd test.Selecting body weight for use is 20 of the Kunming mouses of 18g～22g, male and female half and half.If dosage is 40000mg/kg.Get calculus open capsule 100g adding distil water to 100ml, get in this liquid 24 hours at interval and to irritate stomach 2 times to its mouse oral in 4 hours, irritate at every turn that body of stomach is long-pending to be 0.2ml/10g.Irritate the preceding fasting of stomach 16 hours first.Observed for two weeks record poisoning manifestations and death condition after irritating stomach continuously.
The acute toxicity testing result
The calculus open capsule is to chmice acute toxicity
Figure G2009102240244D0000171
According to above result, after irritating stomach for the Kunming mouse of two kinds of sexes, the calculus open capsule of producing with the Jiangxi Red Star Pharmaceutical of 40000mg/kg dosage do not see obvious poisoning symptom, observe and do not have death in 14 days.Animal subject put to death in the 15th day and dissect main organs such as inspection, liver,spleen,kidney, stomach, intestinal, the heart, lung, show no obvious abnormalities change.To female, male Kunming mouse maximum tolerated dose (MTD) greater than 40000mg/kg.According to acute toxicity grading criteria, belong to nontoxic.
Mouse sperm deformity test: get 25 of the male mice in kunming of body weight 25g～35g, be divided into 5 groups at random, 5 every group, with the positive contrast of the cyclophosphamide of 40mg/kg dosage, the negative contrast of distilled water.3 dosage of test group are respectively 10000mg/kg, 5000mg/kg, 2500mg/kg, get calculus open capsule 50.0g, 25g, 12.5g adding distil water to 100ml respectively, the test solution that is subjected to of preparation corresponding dosage is given mouse stomach (0.20ml/10g), irritate stomach every day once, for three days on end, Yu Weici irritates the 30th day execution animal behind the stomach, gets the epididymis smear, Yihong dyeing, the sperm of 1000 structural integrities of every animal counting calculates distortion spermatogenesis rate, presses x 2Inspection statistics is handled.
The mouse sperm deformity result of the test
The calculus open capsule is to the influence of mouse sperm deformity incidence rate
Group Number of animals (only) Examined sperm count (individual) Sperm deformity number (individual) Rate of teratosperm (%)
High dose ??5 ??5000 ??101 ??2.02
Middle dosage ??5 ??5000 ??98 ??1.96
Low dosage ??5 ??5000 ??108 ??2.16
Negative control ??5 ??5000 ??94 ??1.88
Group Number of animals (only) Examined sperm count (individual) Sperm deformity number (individual) Rate of teratosperm (%)
Positive control ??5 ??5000 ??504 ??10.08
By table as seen, sample does not produce obvious change to the mouse sperm deformity incidence rate, and each dosage group of sample and negative control group comparing difference do not have significance (P＞0.05) and cyclophosphamide positive controls and negative control group comparing difference have significance (P＜0.05) calculus open capsule that mouse sperm is not produced the distortion effect.
30 days feeding trials: the dosage group selection with tried thing and given mode: get 80 of SD rats, male and female half and half, male Mus body weight is 85.31 ± 6.95g, female Mus 83.04 ± 6.83g.Laboratory animal is divided into four groups at random, and promptly matched group and three are tried the thing group, and 20 every group, male and female half and half.If the basic, normal, high dosage of calculus open capsule is respectively 3.00g/kg, 4.50g/kg, 6.00g/kg, basic, normal, high dosage is tried gets calculus open capsule 60g, 90g respectively when thing is prepared, the 120g adding distil water is settled to 200ml, matched group gives the equal-volume distilled water and irritates stomach once every day, irritate the long-pending 1.0ml/100g of being of body of stomach, continuous 30 days.
Experimental technique, all animals of experiment periods give normal diet, single cage is raised, the drinking-water of freely ingesting, observe animal activity and growing state every day, appetite and surplus appetite given in record, claim body weight weekly one time, calculate food-intake weekly, food utilization and total food utilization, test not, fasting 14 hours, hemoglobin is measured in blood sampling, packed cell volume, carry out erythrocyte, leukocyte, platelet count and leukocyte differential count are measured total protein behind the separation of serum, albumin, glutamate pyruvate transaminase, glutamic oxaloacetic transaminase, GOT, blood urea nitrogen, the liver acid anhydride, cholesterol, triglyceride, blood glucose, zootomy is put to death in the blood sampling back, gets liver, kidney, spleen, stomach, duodenum, the section of ovary malingering.
Experimental result, during the nursing in 30 days, each treated animal growth promoter is good, and no abnormal behavior and poisoning symptom do not have death the every index of rat are not produced obvious toxic-side effects.
The pharmacy test of calculus open capsule is as follows
Modern pharmacological research is found, Herba Desmodii Styracifolii can make the litholysis in the urine, its polysaccharide component can suppress water-calcium oxalate crystal and increase, reduce the degree of crystal accumulation, chronic experiment clear-headed rabbit shows, Poria tool diuresis, with ash in contrast, prove that its diuresis is the effect of other compositions beyond the potassium salt, and influence renal tubules Na +Heavily absorption relevant, the ethanol extract of Herba Plantaginis has dose dependent ground and suppresses kidney Na +-K +-atpase activity and produce diuresis, Spora Lygodii can be by increasing ureteral peristalsis and ureter epimere cavity pressure being increased, helping calculus moves down and produces the calculus effect, zoopery shows that Spora Lygodii also has diuresis, points out thus by last medicine to have the calculus diuresis as the calculus open capsule of Main Ingredients and Appearance.
Observe the diuresis and the calculus effect of calculus open capsule by effect experiment
The diuresis experiment
Medicine: calculus open capsule (Jiangxi Red Star pharmaceutcal corporation, Ltd), hydrochlorothiazide tablet (Danyang City, Jiangxi pharmaceutcal corporation, Ltd), ethylene glycol, ammonium chloride all is Guangzhou Chemical Reagent Factory production
Method: 28 of qualified rats are tested in the screening diuresis, body weight 150～200g, the fasting feedwater is divided into 4 groups after one day at random, and each organizes Mus all by 2ml/100g body weight intraperitoneal injection of saline, and gently press abdominal part to make the bladder emptying, positive controls gives 0.05% hydrochlorothiazide aqueous solution, and negative control group waits the normal saline of capacity, and the high dose group low dose group gives 9% and 3% calculus water flowing solution respectively, each group is all irritated stomach by 2ml/100, immediately rat is placed in the metabolic cage after the administration.Compare between each treated animal urine amount of collection and total volume of urine are organized after 1 hour, and take statistics is handled.
The calculus open capsule to the diuresis of mice (X ± S, n=7)
Figure G2009102240244D0000191
Compare 1 with the normal saline group) P＜0.05 2) P＜0.01
Suppress renal calculus and form test
Animal model preparation and medication: 54 of rats, be divided into 3 groups at random, freely drink the water liquid (containing 1.6g ethylene glycol and 0.8g ammonium chloride in every 1000ml aqueous solution) that contains ethylene glycol and ammonification ammonium for every day rat and fed continuously 30 days, to form calculus.Give rat oral gavage 1 time every day, irritating the stomach amount is the 2ml/100g body weight, and experiment is grouped as follows: (1) matched group: normal saline group; (2) low concentration medicine group: 2% calculus water flowing solution; (3) high concentration medicine group: 6% calculus water flowing solution
Detect index and method: rat is put to death in the cervical vertebra dislocation, cuts open the belly and takes out two kidneys, weighs, and vertically cuts kidney open, observes to have or not obvious crystalline deposit thing and free calculus or calcified plaque as judging the calculus standard under anatomic microscope.Divide 5 grades " (-) " according to lithogenous what and kidney shape degree of injury, all visible calculus of renal cortex, medullary substance and renal papillae and crystalline deposit thing, as seen the no abnormal variation of kidney profile " (+) " renal cortex, medullary substance and renal pelvis are dispersed in the crystalline deposit thing on a small quantity; The visible crystalline deposit thing of renal cortex, medullary substance and renal pelvis or little free crystallization " (+++) " is arranged; Kidney portion and visible more crystallization also have free calculus to form, the visible calcified plaque of renal papillae, renal papillae expansion hydrops; " ++ ++ "; Renal cortex, the visible mass crystallization of medullary substance have free calculus to form, the visible calcified plaque of renal papillae, and renal pelvis is obviously expanded hydrops, and the gained enumeration data adopts X 2 test to carry out statistical analysis.
Calculus is logical to the lithogenous influence of kidney of rats
Figure G2009102240244D0000201
The formation rate that found that administration group kidney of rats calculus all significantly reduces, and along with the rising effect of drug level is more obvious.
Experiment shows: 3% calculus water flowing solution (being equivalent to clinical consumption) has tangible diuresis, can make the rat total volume of urine increase about 50%, and acting duration is longer, increase the logical consumption (3 times of amounts) of calculus and only occurring the diuresis peak after the medication between 1～2 hour, total volume of urine and 3% concentration group be there was no significant difference relatively also, may be that body fluid volume is limited, only can strengthen diuretic effect again when acquiring a certain degree by increasing the logical consumption of calculus when urinating.Compare with the hydrochlorothiazide matched group, the logical diuresis of calculus presents gentleness, persistent characteristics, and the formation rate of administration group kidney of rats calculus all significantly reduces in the calculus experiment, and along with the rising effect of drug concentrations is more obvious.
To the cystolithic influence test of the rat opposite sex
Get 60 of male rats, body weight is 200～250g, be divided into 3 groups at random, every Mus all does foreign body vesical calculus moulding: IP injection and defends barbital anesthesia, the sterilization wooden stick of cutting open the belly after will weighing is inserted intravesical and is sewed up the bladder otch, lumbar injection penicillin sodium 80,000 U infection when closing abdomen, postoperative is normally fed.4 weeks beginning ig administration after the type selecting: experiment is grouped as follows, three groups are given and are subjected to reagent thing (production of Jiangxi Red Star Pharmaceutical), negative control group is given JIESHITONG PIAN (production of Guangxi Wuzhou Pharmaceutical limited company), and model control group is given distilled water, once a day, it is 0.60g/kg that dosage is tried three groups, 1.2g/kg, 2.4g/kg, negative control group is 1.2g/kg, the administration volume is 10ml/kg, continuous 30 days.Do not put to death rat next day after time administration, takes out the wooden stick of inserting, and weighs after 60 ℃ of oven dry in 24 hours, deducts original rod and heavily be foreign body in bladder calculus weight and ordinary water group relatively, and seven check between the work group, see Table.
To the cystolithic influence of the rat opposite sex
Group Dosage (g/kg) ??N Calculus weight (mg.x ± s)
Ordinary water ??10ml ??12 ??2.52±0.67
Calculus is logical ??1.2 ??12 ??1.66±0.41*
Be subjected to reagent ??0.6 ??12 ??2.44±0.71
Be subjected to reagent ??1.2 ??12 ??1.69±0.81
Be subjected to reagent ??2.4 ??12 ??1.57±0.32*
Experimental result is found: the weight that heavy dose is subjected to reagent thing group to form calculus is significantly less than the ordinary water group, the difference significance, and prompting this product is brought out the rat bladder calculus to foreign body certain inhibitory action.
Human body lithangiuria (external lithodialysis) test
In 1 branch harness of human body lithangiuria plug test tube with the big or small basically identical of weighing, totally 40 manage, with being subjected to the reagent thing to be mixed with the medicinal liquid of two kinds of concentration, do not prepare JIESHITONG PIAN contrast liquid, and establish the deionized water matched group, the medicinal liquid of every kind of concentration is established 10 pipes, being subjected to reagent liquid concentration is 88.0%, 20.0%, the logical concentration fan 100% of calculus, every pipe by calculus weight add a certain amount of medicinal liquid (g: ml=1: 30) put 37 ℃ of constant temperature water-soluble in, each jolting 1 time (putting upside down several times).Soak and take out calculus after 7 days, weigh after 60 ℃ of 5h oven dry, calculate the lithodialysis rate, soak is got supernatant in semiautomatic biochemistry analyzer mensuration calcium content after centrifugal, and relatively seven checks between the work group of deionized water group:
Group Concentration (%) Weight (mg) before soaking Soak back calcium content (mg) The lithodialysis rate
Be subjected to the reagent thing ??88 ??79 ??1.172 ??2.5
Be subjected to the reagent thing ??20 ??81 ??0.645 ??2.1
Calculus is logical ??100 ??84 ??1.204 ??1.9
Water ??- ??82 ??0.533 ??3.7
Experimental result shows that low concentration is subjected to the lithodialysis rate and the deionized water of reagent thing close, the difference nonsignificance, but the calcium content of high concentration soak is greater than deionized water, the difference significance, and this product has certain litholytic effect to the human body lithangiuria when high concentration.
The influence of calculus open capsule Dichlorodiphenyl Acetate induced mice pain
Get 50 of mices, be divided into 5 groups at random.The administration group is pressed 2.4g/kg, 1.2g/kg, 0.6g/kg dosage ig administration respectively, positive controls is given rotundine, the blank group is given ordinary water, every day, the ig administration was 1 time, the administration volume is 20ml/kg, successive administration 3 days, and the equal ip of each mice of 1n injects 0.7% acetic acid 10ml/kg after the last administration, and write down behind each ip in mice injection acetic acid and to turn round body number and the relatively t check between the work group of ordinary water group in the 15min.
Mice acetic acid is caused the result that influences of pain
Group ??n Dosage g/kg 15 minutes object condition numbers (x ± s) Suppression ratio (%)
Matched group ??10 ??- ??42.9±8.9
The rotundine group ??10 ??80mg ??21.6±6.9 xxx ??49.7
Be subjected to the medicine group ??10 ??2.4 ??27.0±5.9 xxx ??37.0
Be subjected to the medicine group ??10 ??1.2 ??30.7±7.7 xx ??28.4
Be subjected to the medicine group ??10 ??0.6 ??35.9±5.3 x ??15.5
The result as seen, the logical Dichlorodiphenyl Acetate induced mice of calculus is ached significant reaction is arranged.
Mice hot plate piece is caused the influence of pain.
50 of mices are male, be divided into 5 groups at random, first group gives ig injection ordinary water, give rotundine for second group, its excess-three group is pressed 2.4g/kg, 1.2g/kg, 0.6g/kgig injection respectively, and the administration volume is 20ml/kg, measured the pain threshold values of each Mus respectively after the administration in 1,2,3 hour with method, the difference that deducts basic value with value behind the medicine as medicine after the increment of pain valve, with the ordinary water group relatively, do t check between group
The result that influences to pain due to the mice heat
Figure G2009102240244D0000231
Experimental result shows: be subjected to the increment of 3 hours pains of reagent thing heavy dose group administration valve greater than the ordinary water group, and the difference significance, prompting this product has certain analgesia valve action bitterly due to mice heat.
Dimethyl is caused the effect of the ear swelling of mice.
Get 40 of mices, body weight 20g～25g, male and female half and half are divided 4 groups at random, 10 every group, first group for being subjected to reagent 2.4g/kg, second group of reagent 1.2g/kg, the 3rd group of positive contrast aspirin, every day, gastric infusion was 1 time, continuous 3d, after last 1 administration 1 hour, evenly be coated with dimethyl 0.04ml at every mice left side ear and cause inflammation, put to death animal behind the 30min, cut the mice ears along auricle, respectively in the punching of left and right sides auricle same area, the both sides auricle of laying is weighed respectively, calculates swelling value and inhibitory rate of intumesce with the 6mm card punch, the swelling value is checked with t value and t ' value method, and compared between organizing.
Inhibitory rate of intumesce: 1 (%)=(EC-EC)/EC * 100%Et: the average swelling value of administration group, the average swelling rate of Ec matched group
Dimethyl is caused the result of mice ear effect
Group Dosage g/kg Swollen film value (mg) Inhibitory rate of intumesce (%)
Matched group ??20.4±4.7
Be subjected to the medicine group ??2.4 ??14.8±5.3 * ??27.5
Be subjected to the medicine group ??1.2 ??17.6±3.9 ??13.7
Aspirin ??0.2 ??14.5±5.5 * ??28.9
Through comparing, be subjected to heavy dose of 2.4g/kg of reagent thing and aspirin 0.2g/kg all can obviously suppress the mice ear that causes by dimethyl with matched group.
To the effect of mice agar granuloma
Get 40 of 200g ± 20g rat, male and female half and half are divided 4 groups at random, every group 10, under the sterile working, carry out the dorsal line subcutaneous injection, every Mus 2% agar solution 2ml, the gastric infusion that divides into groups then, first group is normal saline, 2nd, 3 groups for being subjected to reagent 2.4g/kg, 1.2g/kg, and the 4th group is aspirin 0.2g/kg, each organizes equal every day of gastric infusion 1 time, and successive administration was put to death animal after 8 days, peel off granuloma, take by weighing the granuloma weight in wet base with electronic scale.The result is with comparing between t value method and the check of t ' value method and Construction Bank's group.
To mice agar granulation effect result of the test
Group Dosage (g/kg) Agar granuloma weight in wet base (g)
Matched group ??1.99±0.32
Be subjected to the reagent group ??2.4 ??1.45±0.56 x
Be subjected to the reagent group ??1.2 ??1.59±0.43 x
Group Dosage (g/kg) Agar granuloma weight in wet base (g)
Be subjected to the reagent group ??0.2 ??14.2±0.50 xx
Compare p＜0.05, p＜0.01 with matched group
Extracorporeal bacteria inhibitor test, tube dilution method is adopted in this test, and experimental result is subjected to the reagent thing to beta hemolytic streptococcus, and the minimum inhibitory concentration of Diplococcus pneumoniae (MIC) is 25mg/ml, to staphylococcus aureus, the minimum inhibitory concentration of bacillus pyocyaneus (MIC) is 50mg/ml.

Claims (8)

1. detection method for the treatment of the components of pharmaceutical composition of diseases of urinary system, the composition of the pharmaceutical composition of wherein said treatment diseases of urinary system is Herba Desmodii Styracifolii, Stigma Maydis, Folium Pyrrosiae, Herba Abri, Poria, Herba Plantaginis, Herba lygodii, Rhizoma Imperatae, and each composition percentage by weight is as follows:
Herba Desmodii Styracifolii 21～24%
Stigma Maydis 12～18%
Folium Pyrrosiae 9～13%
Herba Abri 8～13%
Poria 8～12%;
Herba Plantaginis 7～10%
Herba lygodii 7～10%
Rhizoma Imperatae 8～13%
And chlorogenic acid (C in the described Folium Pyrrosiae 16H 18O 9) content, must not be less than 0.3% of gross weight, above-mentioned medical material is obtained described pharmaceutical composition by following method for making:
Step 1: get described Poria and be divided into two parts, it is 80～90% that a copy of it accounts for the Poria total weight percent, and it is 10～20% that another part accounts for the Poria total weight percent;
Step 2: will account for the Poria total weight percent and be 80～90% portion and be ground into fine powder;
Step 3: will remain Poria and all the other seven flavor medicine materials decoct with water 1～3 time, each 2～4 hours, collecting decoction filtered, and filtrate is condensed into the clear paste that relative density is 1.19～1.20 (90 ℃);
Step 4: add the Poria fine powder, mixing, drying is ground into fine powder, sieves, and adds appropriate amount of auxiliary materials, and mixing is granulated drying, granulate;
Described detection method comprises:
Compositions 5～15g gets it filled, add water 30～70ml and make dissolving, in 60～90 ℃ of extractions 1～3 time, each 30～50ml discards petroleum ether liquid with petroleum ether, ethyl acetate extraction 1～3 time of water liquid, each 20～40ml merges ethyl acetate liquid, evaporate to dryness, residue adds ethanol 1～3ml makes dissolving, as need testing solution;
Other gets Herba Plantaginis control medicinal material 1～3g, adds 70% ethanol, 100～300ml, refluxes 2～3 hours, puts coldly, filters, and filtrate evaporate to dryness, residue add water 30～50ml makes dissolving, extracts 2～4 times for 60～90 ℃ with petroleum ether, gets control medicinal material solution with legal system;
Draw above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, with ethyl acetate-formic acid-water is developing solvent, launches, and takes out, dry, spray is heated to clear spot with the vanillin sulfuric acid solution, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
2. detection method as claimed in claim 1,
The compositions 10g that gets it filled adds water 50ml and makes dissolving, extracts 2 times each 40ml for 60～90 ℃ with petroleum ether, discard petroleum ether liquid, ethyl acetate extraction 2 times of water liquid, each 30ml merges ethyl acetate liquid, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution;
Other gets Herba Plantaginis control medicinal material 1g, adds 70% ethanol 100ml, refluxes 2 hours, puts coldly, filters, and filtrate evaporate to dryness, residue add water 30ml makes dissolving, extracts 2 times with 60～90 ℃ of petroleum ether and rises, and gets control medicinal material solution with legal system;
Draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, with ethyl acetate-formic acid of 6: 1: 1-water is developing solvent, launches, and takes out, dry, spray is heated to clear spot at 105 ℃, in the test sample chromatograph with 5% vanillin sulfuric acid solution, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
3. detection method as claimed in claim 1 also comprises chlorogenic acid (C 16H 18O 9) the step of assay, described assay adopts high effective liquid chromatography for measuring, condition determination is as follows:
Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Acetonitrile-0.5% phosphoric acid solution (11: 89) is a mobile phase; The detection wavelength is 326nm, and number of theoretical plate calculates by the chlorogenic acid peak should be not less than 3000;
It is an amount of that the preparation precision of reference substance solution takes by weighing the chlorogenic acid reference substance, adds methanol and make the solution that every 1ml contains 60 μ g, promptly;
The about 1.0g of this product content under the content uniformity item is got in the preparation of need testing solution, the accurate title, decide, and puts in the 100ml round-bottomed flask, added the 50ml methanol eddy 2 hours, filter, with about 20ml gradation washing medicinal residues of methanol and filter, merging filtrate, evaporate to dryness, residue add methanol make in right amount the dissolving and be transferred in the 10ml measuring bottle, add methanol to scale, shake up, promptly;
Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly.
4. detection method as claimed in claim 1, wherein said Herba Desmodii Styracifolii main component is soybean saponin I, puriri glycosides-1, puriri glycosides-3, Xia Futa basket glycosides, described Stigma Maydis main component is a fatty oil, volatile oil, alkaloid, ascorbic acid, sitosterol, described Folium Pyrrosiae main component is the bracken triterpene, cupreol, kaempferol, Quercetin, isoquercitrin, Trifolin, described Herba Abri main component is a triterpenes: sophoradiol, abrisapogenol A-G. glycyrrhizin, described Poria main component is β-pachyman, the acetyl pachymic acid, pachymic acid, chitin, protein, choline, described Herba Plantaginis main component is an aucubin, Homoplantaginin, ursolic acid, cupreol, described Herba lygodii main component is an aminoacid, saccharide, flavonoid glycoside and phenols, described Rhizoma Imperatae main component is an arundoin, the Yin Baimao element, Isoarborinol alcohol.
5. detection method as claimed in claim 1, amount of water is 10～12 times of amounts of medical material weight in the step 3.
6. detection method as claimed in claim 5, amount of water is 10 times of amounts of medical material weight in the step 3.
7. detection method as claimed in claim 1 decocts with water secondary in the step 3, each 3 hours, collecting decoction filtered, and filtrate is concentrated into the clear paste that relative density is 1.20 (90 ℃).
8. detection method as claimed in claim 1, wherein the optimum weight percentage ratio of each composition is:
Herba Desmodii Styracifolii 22.8%
Stigma Maydis 15.1%
Folium Pyrrosiae 11.4%
Herba Abri 11.4%
Poria 11.4%
Herba Plantaginis 8.25%
Herba lygodii 8.25%
Rhizoma Imperatae 11.4%
And chlorogenic acid (C in the described Folium Pyrrosiae 16H 18O 9) content, must not be less than 1% of gross weight.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110927291A (en) * 2019-12-20 2020-03-27 成都普思生物科技股份有限公司 Detection method of abrus herb extract characteristic spectrum

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110927291A (en) * 2019-12-20 2020-03-27 成都普思生物科技股份有限公司 Detection method of abrus herb extract characteristic spectrum

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Denomination of invention: Method for detecting drug composition components for treating urinary system diseases

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